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鉴于大豆难以获得转基因植株的现实,借鉴Kereszt等建立的发根土壤杆菌(Agrobacterium rhizogenes)K599介导的大豆转化体系,并针对实验中遇到的具体问题,对原体系进行了适当改进。结果表明:改进的体系操作更简单,对发根培养条件、人员操作技能要求更低,所形成的植株更具生命力。同时,应用改进体系以及RNAi技术,构建了表达线虫特异基因dsRNA的转基因发根植株,可在60 d内在转基因大豆发根上对植物线虫基因的RNAi效果完成验证。改进的体系为培育抗大豆胞囊线虫品种奠定了基础。
In view of the fact that it is difficult for soybean to obtain transgenic plants, we used the K599-mediated soybean transformation system of Agrobacterium rhizogenes established by Kereszt et al., And made appropriate improvements to the specific problems encountered in the experiment. The results showed that: the improved system operation is simpler, less demanding on hairy root culture conditions and personnel operation skills, and the formed plants are more viable. At the same time, RNAi technology and improved system were used to construct transgenic hairy root plants expressing nematode-specific gene dsRNA, which verified the RNAi effect of plant nematode gene on transgenic soybean hairy roots within 60 days. The improved system laid the foundation for cultivating anti-soybean cyst nematode.